Medical Uses of Glucans

ABSTRACT

The invention relates to a glucan having a beta-(1,3)-backbone with one or more beta-(1,3)-side chains linked thereto for use in the treatment of asthma and related diseases of abnormal pulmonary function in an animal. Also described is a method of treating asthma and related diseases of abnormal pulmonary function in an animal comprising administering to said animal an effective amount of a glucan having a beta-(1,3)-backbone with one or more beta-(1,3)-side chains linked thereto.

The present invention relates to the use of glucans for treating asthmaand related diseases of abnormal pulmonary function.

Glucans are a heterogeneous group of glucose polymers found in the cellwalls of plants, bacteria and fungi. The basic structural unit inbeta-glucans is a backbone chain comprising or consisting ofβ(1→3)-linked glucosyl units. Depending upon the source and method ofisolation, beta-glucans have various degrees of branching and oflinkages in the side chains. The frequency and type of linkage in theside chains determine the molecule's biological activity. Beta-glucansof fungal and yeast origin are normally insoluble in water, but can bemade soluble either by acid hydrolysis or by derivatization introducingforeign groups like -phosphate, -sulphate, -amine, -carboxymethyl and soforth to the molecule.

In Europe, Asia and USA, beta-glucans especially from Bakers' yeast havelong been employed as feed additives for animals, as dietary supplementfor humans, in treatment of wounds, and as an active ingredient in skincream formulations. Further, glucans have been employed as functionalpharmaceutical agents exemplified by their application for treatment ofcancer as shown in WO02058711. Different glucans have very differentactivities in vivo and it is usually hard to predict which glucan wouldbe most effective for a given therapy and not possible to assume that aparticular glucan will be active just because a glucan with a differentstructure has been shown to have that activity.

Asthma is a chronic inflammatory lung disease characterized by dyspnoea,superimposed infections, mild to grave impairment of general health, insome cases leading to a painful death. The prevalence of the disease hasincreased markedly over the last few decades such that about one in tenpersons in industrialized countries carry the diagnosis. The efficacy ofavailable therapy is modest and there is no known effective prophylaxis.The impact of the disease on public health is therefore serious, relatedhealth care expenditures enormous and increasing.

The observation that the prevalence of asthma has increased dramaticallyover the past two decades while certain environmental exposures (e.g.exposure to infectious diseases) have decreased, suggests that certaininfections may reduce the risk of developing asthma, presumably byenhancing the development of protective immunity. These observations arethe basis for the ‘hygiene hypothesis’, which suggests that improvedhygiene in industrialized societies, which improved public healthmeasures and the use of vaccines and antibiotics, has reduced theincidence of infections and the exposure to environmental factors thatwould normally stimulate the immune system in a way that mitigatesagainst asthma.

At present asthma is not cured, it is managed, through careful controlof the patient's environment, measurements of lung function toanticipate an attack and through medication. Medication includesanti-inflammatory drugs which reduce the spontaneous spasm of themuscles of the airways and bronchodilators which increase the diameterof the air passages and thereby the flow of gases to and from the lungs.

Asthma is caused by dysregulated immune responses in the respiratorymucosa. It is believed that Th2(T helper 2)-driven responses play acrucial role in the pathogenesis, particularly because typical Th2cytokines are involved: interleukin 4 (IL4) and IL-13 enhance IgEproduction, IL-4, IL-9 and IL-10 enhance mast cell activity, IL-5enhance eosinophil accumulation and IL-9 and IL-13 directly enhancemucus secretion and airway hyperactivity—all of these phenomena typicalof asthma.

Although it is generally accepted that Th2-driven immune reactions areinvolved in the development of asthma and related diseases of abnormalpulmonary function, the exact pathogenesis is not understood, nor arethe protective mechanisms in non-asthmatic individuals. Although allindividuals are exposed to allergens and other sensitizing agents in theenvironment, asthma develops only in some of the exposed, suggestingthat both environmental and genetic factors play important roles indetermining the development of pathogenic Th2 responses.

JP 2002335926 suggests the use of a glucan containing formulation fortreating a wide variety of different conditions including HIV, diabetesand asthma. The glucans here are unbranched or have 1,6-linked single ormultiple glucopyranose units and do not have 1,3-linked side chains.

Surprisingly, the inventors of the present invention found that asthmaand related diseases of abnormal pulmonary function might be treated byusing a certain class of glucan.

Even though extensive studies have been performed on asthma and relateddiseases and conditions, no single effective and patient-compliantmethod and substance has been found for the treatment and/or preventionof these illnesses. The present invention addresses this problem,proposing the use of certain glucans for preventing and/or treatingasthma and related pulmonary diseases.

Thus, in one aspect, the invention provides the use of a glucan having abeta-1,3 backbone with one or more beta-1,3 side chains linked thereto,in the manufacture of a medicament for the treatment of asthma orrelated diseases of abnormal pulmonary function.

Alternatively viewed, the invention provides a glucan having a beta-1,3backbone with one or more beta-1,3 side chains linked thereto for use inthe treatment of asthma or related diseases of abnormal pulmonaryfunction.

The glucan has a beta-1,3 backbone, i.e. the backbone is made up ofbeta-1,3 linked glucopyranose units. The glucan has beta-1,3 sidechains, i.e. attached to the backbone via a beta-1,6 linkage are one ormore side chains made up of beta-1,3 linked glucopyranose units. Thusthe side chain comprises 2 or more, typically 3, 4, 5, 10 or morebeta-1,3 linked glucopyranose units.

Thus, preferably less than 10%, more preferably less than 5%, mostpreferably less than 3% or 2% of the glycosidic bonds in the glucanmolecule will be beta-(1,6)-linkages.

Preferably the glucan moiety is underivatised by chemical groups, atleast, it preferably does not contain the following groups: sulfate,acetate, phosphate or phosphonate.

In the present invention the glucan is administered to a subject by anypossible mode of administration, but preferably orally.

The medicament may be administered as part of a dietary regimen. Themedicament may be formulated as a nutraceutical, animal feed, food, partof a nutraceutical, animal feed or food and/or adjuvant. The glucancontaining medicaments may be administered to any animal, includinghumans, non-human primates and other mammals, domestic and livestockanimals, birds, and fish, including farmed birds and companion birdslike parrots, and farmed fish and pet fish. Specific examples includedog, cat, horse, cow, pig, goat, rat, mouse and sheep. Humans arepreferred recipients.

The glucan of use according to the invention is not part of a naturalwhole product, e.g. a plant which may have within it protein and glucan.Thus the glucan can be considered a processed product. It may beconsidered isolated in the sense that it is no longer part of the wholecell or organism from which it originates, preferably it is no longer inadmixture with all the other cell wall components which it is found within nature. Preferably the glucan is also processed in the sense that thechemical structure is altered as compared to the naturally occurringstructure as well as being isolated (at least partially) from othernaturally occurring cell-wall components. The alterations will typicallycomprise reductions in length of the backbone and/or in length orcomplexity of branches.

The glucan can be from a variety of different sources. Important andpreferred sources are yeasts as exemplified by Saccharomyces cerevisiae.The yeast cell wall consists mainly of polysaccharides made up of threesugars, glucose, mannose, and N-acetylglucosamine. The mannosepolysaccharides are linked to proteins to form a mannoprotein layermainly localized at the external surface.

Glucans from yeast consist essentially of a beta-1,3 backbone and sidechains which are generally linked via a beta-1,6-linkage to the backbonestructure. Therefore beta-glucans from yeast and other organismsproducing glucans with a similar structure are often described asbeta-1,3/1,6; beta-1,3 or simply beta-glucans.

Preferred beta-glucan containing products for use according to theinvention contain at least 75%, preferably at least 80%, carbohydrate asa percentage of total cell components. Of this carbohydrate, themajority is glucan.

Examples of useful beta-glucan products for the present inventioninclude, but are not limited to, the glucan products Imucell asmanufactured by Biothera and Immiflex (formerly Fluflex) as distributedby CarePharma Co, Ltd.

Particularly preferred beta-glucans include, but are not limited to,particulate and soluble yeast cell wall glucans as described inPCT/IB95/00265 and EP 0759089. Depending upon yeast strain and type,glucan constitutes up to 25% of the yeast cell wall dry weight. Duringthe process of isolating beta-glucan from yeast the outer layer ofmannoprotein is removed as well as most of the inner content of thecell, leaving a “ghost” particle, or whole glucan particle, constitutingthe beta-glucan layer. If the beta-glucan is isolated from autolysedyeast, the cell wall is more collapsed giving a crumpled ghost particle.

Brewers yeast differs in composition from bakers yeast because it isgrown under anoxic conditions, resulting in a lower level of beta-glucanin the cell walls. Other yeasts which provide a source for the glucaninclude Candida sp., Hansenula sp., Histoplasma sp., Kloeckera sp.,Kluyveromyces sp., Pichia sp., Rhodotorula sp., Saccharomyces sp.,Schizophyllum sp., Schizosaccharomyces sp., Torula sp. and Torulopsissp.

As discussed above, surprising efficacy has been shown for a glucanhaving a beta-1,3 backbone with one or more beta-1,3 side chains linkedthereto. Such glucans may also be sourced from some of the followingknown glucan sources.

Other sources of glucan are mushrooms or other fungi exemplified bythose belonging to the classes Mastigomycotina, Ascomycotina,Basidiomvcotina, and Deuteromycotina (imperfect fungi). Other suitablefungi include Aspergillus sp., Penicillium sp., Sclerotinia sp., andSclerotum sp.

A third source of glucan are the members of the Gramineae (grasses),amongst the Angiosperms, where they are major components of endospermwalls of commercially important cereals such as oat, barley, rye,sorghum, rice and wheat. Apart from these, plants are not preferredsources.

A fourth source of glucan are algae, exemplified by the classesChlorophyceae, Charophyceae Euglenophyceae, Phaeophyceae,Bacillariophyceae, Chrysophyceae, Xanthophyceae, Pyrrophyceae andRhodophyceae. Laminarin is an example for a glucan product fromsea-weed.

It is also possible to derive said glucan from the cell walls ofBacteria like Alcaligenes(Achromobacteriaceae), Agrobacterium andRhizobium(Rhizobacteriaceae). Examples are Alcaligenes faecalis,Agrobacterium radiobacter and A. rhizogenes, Rhizobium japonicum og R.trifolii, Streptococcus pneumoniae as well as the CyanobacteriaceaeAnabaena cylindrica. Finally, also the glucan from Streptococcusfeacalis which is the origin for the glucan Curdlan may be used inconnection with the present invention.

In the animal feed and farming industry the cells of organisms, mostoften yeast cells, are used, and fed directly to the animals. Theseproducts come in different forms and shapes, like compressed, liquid,crumbled, dry, active, in-active cells, and combinations like activedry, instant active dry and inactive dry. These products are most oftenthe remnants of the cells used for other production processes likebrewing or baking.

Likewise, beta-glucan formulations solublized by derivatization, e.g.,glucan amines or carboxymethyl-glucans are possible active products.

It is convenient to use processed products or cell extracts to achievethe effect of the present invention. These products may be hydrolysed orautolysed cells, partially or completely purified cell walls. All theseproducts are available in various forms suited to different types ofuse: liquid, semi-paste, paste, fine powder, oil-coated powder,micro-granulated powder, to mention only some.

Products containing isolated carbohydrate components may be combinationproducts of two or more components (e.g. from the yeast cell wall), forexample a combination of glucans and mannans.

The glucan may be mixed with other components e.g. other parts of thecell wall such as mannans or components not being part of the cellwalls, like vitamins or minerals and other agents frequently used in thepharmaceutical, nutraceutical, food, animal feed and veterinaryindustry. Examples of this group of products are ready to useglucan-products combined with minerals and vitamins as well asnutraceuticals combining glucans and other anti-asthmatic agents.

In addition to the 1,3 linked side chains, the glucan may also have oneor more 1,6 linked side chains. However, preferred glucans are thosewhich have been treated by acid or enzyme or any other suitable methodto significantly reduce or eliminate the number of repetitive(1,6)-linked glucose molecules within the glucan, or occur naturallywith low levels of 1,6 linkages. These (1,6)-linked glucose moieties aremainly in a beta-conformation, and would normally be found in the sidechains of the beta-glucan molecule. The number of beta-1,6-linkedglucose moieties can vary from one to a significant proportion of theglucose moieties depending on the source of glucan. The resultingpreferred glucans have beta-1,3-main chains and beta-1,3 side chainswhich are linked together by a single beta-1,6-linkage which is notcleaved off by the elimination treatment. These modified glucanmolecules are preferably derived from S. cerevisiae.

The preferred glucans are essentially free of repetitive beta 1,6-linkedglucosyl units. Thus, the 1,6-linkages at the branch points do notprovide ‘repetitive’ beta 1,6-linked glucosyl units. By ‘essentiallyfree’ is meant less than 2%, preferably less than 1% of the totalglucosyl units. An example of such a product is seen in FIG. 1 being a¹H-NMR-spectrum of a branched beta-1,3-glucan with <1% repetitivebeta-1,6-linked glucosyl units.

Some treatments, such as enzyme treatments, may leave up to 4beta-1,6-linked glucosyl units uncleaved in the side chains. Suchmolecules are also ‘essentially free’ of repetitive beta 1,6-linkedglucosyl units.

The glucan which can be used in relation to the present invention couldbe in the form of a single, extracted fraction or two or more differentfractions with different molecular weights.

The most preferred source for the glucan for the present application arecell walls from Saccharomyces cerevisiae. Of these, a preferred sourcefor use in the present invention is the soluble yeast product SBG(Soluble Beta Glucan) as produced by Biotec Pharmacon ASA, a Norwegianbased company.

The product is an underivatized (in terms of chemical modifying groups)aqueous soluble beta-1,3/1,6-glucan, characterised by NMR and chemicalanalysis to consist of polymers of beta-1,3-linked D-glucose containingside-chains of beta-1,3 and beta-1,6-linked D-glucose, wherein thenumber of beta-1,6 moieties in the side chains (not including at thebackbone/side chain branch point) is considerably reduced as compared tothe structure of said glucan in the yeast cell wall. An example of sucha composition is as follows:

COMPOSITION Value/range typical value WATER 977-983 gram/kg 980CARBOHYDRATES 18-22 gram/kg 20 PROTEINS max 1 gram/kg <1 ASH max 1gram/kg <1 LIPID Max 1 gram/kg <1

The molecular structure of SBG is as follows:

n≧0; R=H or (C₆H₈₋₁₀O₅)_(m); m (R1+R2)=35 to 2000 glucosyl units.

The reduction of the beta-(1,6)-linked glucosyl residues to produce theabove preferred glucan of the present invention may be achieved in oneof the following ways:

i) Enzymatic treatment, for example as described in Norwegian Patent No.300692:

The side chains of beta-1,6-linked glucose in the micro-particulateproduct prepared as in U.S. Pat. No. 5,401,727 are selectively removedby enzyme treatment with an enzyme which specifically acts onbeta-1,6-linkages in a poly-glucose chain. The micro-particulate product(0.2 grams) is suspended in 40 ml 50 mM ammonium acetate buffer at pH5.0 and mixed with 20 units of the beta-1,6-glucanase enzyme. Themixture is continuously stirred for 6 hours at 37 degrees Celsius andthe action of the enzyme stopped by boiling for 5 minutes. The residualenzyme treated particles are washed repeatedly in sterile distilledwater by centrifugation and re-suspension. The resulting product is abranched beta-1,3-glucan with beta-1,3-glucan side chains connected bybeta-1,6-linked at the branching points, and being essentially free ofbeta-1,6-linked glucose in the side chains which extend from thebranching points. The key step being incubation of a particulate glucanwith a beta-1,6-glucanase enzyme at 32 to 40° C. for 3 to 9 hours.

ii) Formic Acid treatment: For example, a micro-particulate productprepared as in U.S. Pat. No. 5,401,727 may be suspended in formic acidand heated. The suspension is cooled and free formic acid removed.

The soluble glucans of the present invention have a molecular weight ofgreater than 6000 Da, preferably in the range of about 6 kDa to 30 kDA,more preferably in the range from about 10 kDa to 25 kDa, mostpreferably in the range of about 15 kDa to 20 kDa. When in aqueoussolution the molecules may take part in interchain interactions giving ahigh molecular weight appearance of up to 5000 kDa when analysed by gelperformance chromatography.

A preferred glucan containing formulation for use in the invention is amixture of soluble beta-glucan molecules with molecular weights(MW)>6000 daltons that interact to give a higher order conformation. Forexample, a mixture of linear beta-1,3-glucan chains with a numericalMW>6 kDa, preferably with a MW ranging from 6-30 kDa, together withbranched high molecular weight beta-1,3-glucans (e.g. MW>15 kDa) withbeta-1,3 linked side chain(s) extending from within the main chain asshown above.

Preferably, the glucans have an average molecular weight of about 15-20kDa, with a range from about 6 to about 30 kDa, preferably from about 10to about 25 kDa. When in aqueous solution the molecules may take part ininterchain interactions giving a high molecular weight appearance of upto 5000 kDa when analysed by gel performance chromatography. Preferredcompositions are those that form a gel like appearance in aqueoussolution, demonstrating complex intermolecular interactions.

Yet another preferred product for use in connection with the presentinvention is NBG (Norwegian Beta Glucan), a particulate yeast product asproduced by Biotec Pharmacon ASA. NBG is a product derived from BakersYeast (Saccharomyces cerevisiae). The product is a natural underivatized(in terms of chemical modifying groups) particulate beta-1,3/1,6-glucan,characterised by NMR and chemical analysis to consist of polymers ofbeta-1,3-linked D-glucose containing side-chains of beta-1,3 andbeta-1,6-linked D-glucose.

Typical values for the chemical composition of NBG are as follows:

COMPOSITION % by weight Typical range CARBOHYDRATES Min 75 75-80 LIPIDSMax 5 3-5 NITROGEN Max 1.4 0.8-1.2 ASH Max 12  8-10 TOTAL SOLID Min 9595-98

The basic common molecular structure of SBG and NBG, which representspreferred beta-glucans for use in the present invention, is as follows:

R=H or (C₆H₈₋₁₀O₅)₁₋₅₀; n=35-2000;

The preferred particulate beta-glucan of the present invention may beprepared in the following way:

By repeated extractions in alkali and acid of dry Saccharomycescerevisiae, for example according to the procedure described in U.S.Pat. No. 5,401,727 (incorporated herein by reference). The extractionprocess described removes cytoplasmic components inside the yeast cellsas well as the mannose containing polysaccharides and proteoglycanswhich are on the cell surface. The product prepared according to thisprocedure consists of a beta-1,3 beta-1,6-glucan with a particle size of2-5 micrometers. The chemical structure of this micro-particulatebeta-1,3 beta-1,6-glucan is characterized by 83% beta-1,3 linkedglucose, 6% beta-1,6 linked and 5% beta-1,3,6 linked glucose, and it isa beta-1,3-glucan chain with beta-1,3,6-linked glucose as the branchpoints.

The particulate glucans of the present invention have a molecular weightin the range of 5000 Da to 1,000,000 Da, preferably in the range of 25kDa to 500 kDa, more preferably in the range of 150 kDa to 300 kDa andmost preferably about 250 kDa.

The particulate glucans described above may be solubilized as describedin WO/2001/062283 (incorporated herein by reference). Thus, formic acidcan be used to both reduce the number of beta-(1,6)-linked glucosylresidues in the glucan and to solubilize the glucan.

Other structures and/or structural conformations within the family ofbeta-1,3-glucans can be readily identified or isolated by a person ofordinary skill in the art following the teaching of this invention. Theabove is thus a guideline to achieve a highly potent product, but is nota limitation towards even more potent products. Isolated structuralelements of the complex mixture may have improved effects over thepresently exemplified formulations when administered.

The beta-glucans used in accordance with the present invention haveutility as safe, effective, therapeutic and/or prophylactic agents,either alone or as adjuvants, to enhance the immune response in humansand animals by inducing a local inflammatory response by stimulating orpriming the systemic immune system to release certain biochemicalmediators (e.g., IL-1, IL-3, IL-6, IL-17, TNF-α, and GM-CSF). Thisspecific effect is unique to these beta-glucans while many similarglucans claim not to stimulate or prime the immune system in thatmanner.

Without wanting to be bound by theory, it is believed that thebeta-glucans used in accordance with the present invention modulate theimmune system in humans and animals by down-regulating the Th2 immuneresponse and by up-regulating the Th1 immune response.

Suitable carriers or auxiliaries for use in formulating glucancontaining compositions for use in the present invention includemagnesium carbonate, titanium dioxide, lactose, mannitol and othersugars, talc, milk protein, gelatin, starch, vitamins, cellulose and itsderivatives, animal and vegetable oils, polyethylene glycols andsolvents, such as sterile water, alcohols, glycerol and polyhydricalcohols. The pH and exact concentration of the various components ofthe composition are adjusted according to routine skills.

The compositions for medical and veterinary use are preferably preparedand administered in dose units. The term “dose units” and itsgrammatical equivalents as used herein refer to physically discreteunits suitable as unitary dosages for the human or non-human subject,each unit containing a predetermined effective amount of glucancalculated to produce the desired therapeutic effect in association withthe required physiologically tolerable carrier, e.g., a diluent or avehicle.

The composition may comprise the active ingredient alone, in a formsuitable for administration to a subject, or the composition willtypically comprise the glucan and one or more physiologically acceptablecarriers, one or more additional active ingredients, or some combinationof these.

The formulations described herein may be prepared by any method known orhereafter developed in the art of pharmacology, veterinary science,animal and human nutrition etc. In general, such preparatory methodsinclude the step of bringing the active ingredient into association witha carrier or one or more other accessory ingredients, and then, ifnecessary or desirable, shaping or packaging the product into a desiredsingle or multi-dose unit. Controlled or sustained-release formulationsof a composition of the present invention may be made using conventionaltechnology.

Dosage levels of the active compounds comprised in the composition foruse in the present invention may vary. Functional dose ranges of theglucans can be readily determined by one of ordinary skill in the art.For example, when administered orally the functional dose range andeffective amount for a human would be in the region of 10-500 mg/kg b.w.(body weight)/day, preferably 50-300 mg/kg b.w./day. When administeredparenterally a suitable functional dose range would be 0.1-10 mg/kgb.w./day.

The compositions according to the invention may be presented in the formof an article or carrier such as a tablet, coated tablet, lozenges,troches, syrups or elixirs, liposomes, powder/talc or other solid,solution, emulsions, suspension, liquid, spray, gel, drops, aerosol,douche, ointment, foam, cream, gel, paste, microcapsules, controlledrelease formulation, sustained release formulation or any other articleor carrier which may possible or useful in light of the intended orpreferred mode of administration.

The route(s) of administration will be readily apparent to the skilledartisan and will depend upon any number of factors including the typeand severity of the disease being treated, the type and age of thesubject being treated, and the like. The most preferred route ofadministration is orally, optionally by gavage. Formulations suitablefor oral administration of the glucan (preferably soluble glucan)include, but are not limited to, an aqueous or oily suspension, anaqueous or oily solution, or an emulsion. Such formulations can beadministered by any means including, but not limited to, soft gelatincapsules. Liquid formulations of a pharmaceutical composition of thepresent invention which are suitable for oral administration may beprepared, packaged, and sold either in liquid form or in the form of adry product intended for reconstitution with water or other suitablevehicle prior to use. With regard to the particulate product, it ispossible to use other means of administration including but not limitedto capsules, tablets, powders, granules, lozenges, drops, suppositoriesor any other means of administration suitable for a particulate product.Other types of administration which are preferred are nasaladministration, inhalation or any other easy-to-use application in caseof an asthmatic attack. Whatever route is selected, preferably asystemic effect is achieved and the dose and method of administration isselected or designed with this objective in mind.

Therapy may be repeated intermittently while the symptoms are present oreven when they are not present. It might be relevant to administer thecomponents two weeks prior to the expected challenge and/or for severalweeks after the challenge. Continuous use is also possible, as for thetreatment of chronic conditions.

The glucan may be provided alone or in combination with othermedicaments to provide an operative combination. Thus in a furtherembodiment is provided a product containing (a) a glucan as describedabove, and (b) a second active agent for the treatment of asthma orrelated diseases of abnormal pulmonary function, as a combinedpreparation for simultaneous, separate or sequential use in thetreatment of asthma or related diseases of abnormal pulmonary function.

Thus, it is possible to use a single glucan, a combination of two ormore glucans or, if applicable, a combination of glucan(s) and anothermedical substance. With regard to a composition including two or moreglucans it is possible to use different glucans from the same ordifferent species or from the same species but produced by differentmethods.

A skilled artisan/physician will be able to select the medicalsubstances which can be applied together with glucans for treatment ofthe relevant condition. Examples of suitable additional medicalsubstances are, but are not limited to, relief medication likebronchodilators, short-acting, selective beta2-adrenoceptor agonists,such as salbutamol (albuterol USAN), levalbuterol, terbutaline andbitolterol, adrenergic agonists such as inhaled epinephrine andephedrine tablets, anticholinergic medications, such as ipratropiumbromide, chromoglycate therapy; prevention medication like inhaledcorticosteroid, inhaled glucocorticoids including ciclesonide,beclomethasone, budesonide, flunisolide, fluticasone, mometasone, andtriamcinolone. Leukotriene modifiers (montelukast, zafirlukast,pranlukast, and zileuton). Mast cell stabilizers (cromoglicate(cromolyn), and nedocromil), Antimuscarinics/anticholinergics(ipratropium, oxitropium, and tiotropium), Methylxanthines (theophyllineand aminophylline), Antihistamines, Omalizumab, Methotrexate; as well asalternative and different medication used in complementary medicine.

In general, the beta-glucan can be administered to an animal asfrequently as several times daily, or it may be administered lessfrequently, such as once a day. The treatment will for instance dependupon the type of asthma or related disease, the severity of thecondition, and the condition of each patient. The glucan treatment maybe closely interrelated with any other treatment regimen, and could beahead of, concurrent with, or after the administration of any othermedicament.

The glucan or compositions of two or more glucans as described in thepresent invention may be applied as prophylaxis for prevention ofasthmatic conditions in advance of an asthmatic attack or as a treatmentafter the asthmatic incidence has occurred. Thus, ‘treatment’ as usedherein includes prophylactic treatment, i.e. prevention, both of anindividual episode or attack and of the onset of the disease in asubject deemed at risk. Prophylactic treatment will usually be inrelation to an attack/episode. ‘Treatment’ includes a measurable andbeneficial improvement in one or more, preferably more than one symptomof or risk factor for asthma or a related disease of abnormal pulmonaryfunction. Preferably in one or more symptoms and more preferably aconclusion by the patient and/or treating physician that the asthma orrelated disease is improved, either in terms of the historicalpresentation of the disease or what was anticipated (e.g. in the case ofa prophylactic treatment).

The term “asthma” refers to a chronic or recurring inflammatorycondition of the respiratory system in which the airway developsincreased responsiveness to various stimuli, characterized by bronchialhyper-responsiveness, inflammation, increased mucus production, andintermittent airway obstruction, often in response to one or moretriggers. These acute episodes may be triggered by such things asexposure to an environmental stimulant (or allergen), cold air, exerciseor exertion, or emotional stress. Occupational asthma is included. Thesymptoms of asthma can range from mild to life threatening. Treatmentwill typically include an improvement in one or more of the abovesymptoms, by in bronchial inflammation or mucus production as well as atendency towards more normal respiratory behavior in general.

Most advantageously the invention provides for an improvement inbronchial asthma, specifically with regard to a reduction in pulmonaryinflammation and/or a reduction in airway airflow resistance. TheExamples provide protocols for measurement of these parameters.

‘Related diseases of abnormal pulmonary function’ include those diseaseswhere patients exhibit continuous or sporadic impaired pulmonaryfunction, generally associated with restricted airflow and/or bronchialinflammation. Such related diseases include lung disease, exemplified,but not limited to, allergic bronchopulmonary aspergillosis, (chronic)bronchitis with an allergic element and emphysema.

The present invention also provides a method of treating asthma or arelated disease of abnormal pulmonary function in a subject whichcomprises administrating an effective amount of a glucan as describedabove. Typically the subject will be an animal, preferably a human, whohas been diagnosed with or being at risk of asthma (or said relateddisease).

This invention also provides a kit or an administration devicecomprising a glucan as described herein and information material whichdescribes administering the glucan to a human or other animal. The kitor administration device may have a compartment containing the glucan.As used herein, the “Information material” includes, but is not limitedto, a publication, a recording, a diagram, or any other medium ofexpression which can be used to communicate the usefulness of thecomposition of the invention for its designated use. Examples ofadministration devices are, but are not limited to, all types ofinhalers, nebulisers and dry-powder devices.

Thus, in a further preferred embodiment, the invention provides aninhaler, nebuliser or dry-powder device containing a glucan as describedherein in a form suitable for self-administration by a patient to treator prevent an asthma attack. Such a device is typically designed for useby mouth. Thus, preferred devices have the glucan in solubilized liquidor dry powder form. Liquids may be under pressure and the device mayhave components which allow air or oxygen to pass, through apredetermined amount of the glucan making it available for inhalation bythe patient.

By “an effective amount” is meant an amount of a compound effective toameliorate the symptoms of, or ameliorate, treat, prevent, delay theonset of or inhibit the progression of a disease. Ultimately, theattending physician or veterinarian will decide the appropriate amountand dosage regimen. The “effective amount” of the active ingredientsthat may be combined with the carrier materials to produce a singledosage will vary depending upon the subject treated and the particularmode of administration.

Various documents including, for example, publications and patents, arerecited throughout this disclosure. All such documents are, in relevantpart, hereby incorporated by reference. The citation of any givendocument is not to be construed as an admission that it is prior artwith respect to the present invention. To the extent that any meaning ordefinition of a term in this written document conflicts with any meaningor definition of the term in a document incorporated by reference, themeaning or definition assigned to the term in this written documentshall govern.

Referenced herein are trade names for components including variousingredients utilized in the present invention. The inventors herein donot intend to be limited by materials under a certain trade name.Equivalent materials (e.g., those obtained from a different source undera different name or reference number) to those referenced by trade namemay be substituted and utilized in the descriptions herein.

The compositions described herein may comprise, consist essentially of,or consist of any of the elements as described herein.

In a further aspect the present invention provides for the use of theglucans and formulations described above in the manufacture of amedicament for treating autoimmune diseases which are characterized bythe body's immune responses being directed against its own tissues,causing prolonged inflammation and subsequent tissue destruction. Suchdiseases comprise all diseases where a shift of Th1/Th2 reactionpatterns towards a more pronounced Th2-pattern is an essential elementof the pathogenesis. Examples for these autoimmune diseases are, but arenot limited to, celiac disease, Crohn's disease, pancreatitis, systemiclupus erythematosus, Sjogren's syndrome, Hashimoto's thyroiditis, andother endocrinopathies. Multiple sclerosis is also a part of this group.Equivalent methods of medical treatment are also provided.

In a preferred further aspect the present invention provides for the useof the glucans and formulations described above in the manufacture of amedicament for treating allergic rhinitis (hay fever). Equivalentmethods of medical treatment are also provided. The efficacy of theglucans in this therapeutic context is demonstrated in the Exampleshereto.

In a further aspect the present invention provides for the use of theglucans and formulations described above in the manufacture of amedicament for treating allergies. Allergies include mold spores, drugs,foods (in particular nuts, sesame, seafood, egg, soy, milk, wheat andmaize), insect stings, latex and animal products (especially animal hairand dander, cockroach calyx and dust mite excretion. Equivalent methodsof medical treatment are also provided.

The following examples are intended to be illustrative of the presentinvention and to teach one of ordinary skill in the art to make and usethe invention. These examples are not intended to limit the invention inany way. The invention will now be further described in the followingExamples and the figures in which:

FIG. 1 is an ¹H-NMR-spectrum of a branched beta-1,3-glucan with <1%repetitive beta-1,6-linked glucosyl units. The different observedchemical shifts are represented in Table 1 below:

TABLE 1 Chemical shift (ppm) Assignment Comment 5.00 H1 RT(α) H1 in theα-anomer for the reducing terminus (RT) 4.54 H1 BC H1 in backbone chainof (1- 3)-linked glucosyl repeat units (GRUs) 4.39 H1 NRT + H1 H1 in thenon-reducing RT(β) terminus (NRT) + in the β- anomer of the reducingterminus (RT) 4.27 H1 (1-6) SC H1 in (1-6)-linked side- chains 4.03 H6(1-6) SC H6 in (1-6)-linked side- chains 3.72 H6 BC H6 in the backbonechain of (1-3)-linked glucosyl repeat units (GRUs) 3.48 H3 BC + H6′ BCH3 and H6′ in the backbone chain of (1-3)-linked glucosyl repeat units(GRUs) 3.30-3.24 H2 BC + H4 BC + H2, H4 and H5 in the backbone H5 BCchain of (1-3)-linked glucosyl repeat units (GRUs) 3.09 H2 NRT H2 in thenon-reducing terminus (NRT) 3.02 H2 (1-6) SC H2 in (1-6) linked side-chains 2.54 DMSO The solvent peak

FIG. 2 is a graph showing the influence of glucan (BBG) on nasal mucosa.

FIG. 3 is a graph showing the influence of glucan (BBG) on IgE levels inblood over time.

FIG. 4 is a graph showing the effect of glucan (BBG) on intracutaneousreaction over time.

FIG. 5 is a graph showing the effect of glucan (NBG) administration onthe condition of the nasal mucosa.

FIG. 6 is a graph showing the effect of glucan (NBG) administration onthe frequency of sneezing.

FIG. 7 is a graph showing the effect of glucan (NBG) administration oneye itching.

FIG. 8 is a graph showing the effect of glucan (NBG) administration onthe nasal breathing ability.

FIG. 9 is a graph showing the total effect of (NBG) glucanadministration on all four symptoms of hay fever combined. In additionthe pollen presence in that specific time period was obtained andincluded in the figure.

FIG. 10 is a chart showing the results of a questionnaire on all 9subjective symptoms. (9% no effect, 49% effect, 42% uncertain.

FIG. 11 is a graph showing the effect of glucan (SBG and NBG) treatmentof five week-old AKR mice on the production of various cytokines ascompared to sham treated and naïve mice as determined by ELISA.

FIG. 12 is a graph showing the effect of glucan (SEG and NBG) treatmentof five week-old AKR mice on the lung mRNA levels of various cytokinesas compared to sham treated and naïve mice as determined by quantitativereal-time PCR.

FIG. 13 is a graph showing the effect of glucan (SBG and NBG) treatmentof five week-old AKR mice on OVA-specific serum IgE, IgG1 and IgG2alevels as compared to sham treated and naïve mice as determined byELISA.

EXAMPLES Example 1

Five-week-old AKR mice were pre-treated with oral β-glucan daily for 5consecutive days weekly, for 4 Weeks prior to sensitization—particulate1 mg, particulate 6 mg, water-soluble 1 mg, or water-soluble 6 mg. Theycontinued treatment throughout the sensitization and challenge phase.Sham treated mice received an equivalent volume of water. Naïve miceserved as negative controls. β-glucan treated mice and sham treated micewere then intraperitoneally sensitized to ovalbumin (OVA) and aluminiumhydroxide twice at a 7 day interval, followed by 4 sets of intratrachealchallenges at 7-10 day intervals with OVA. Bronchoalveolar lavage fluid(BALF) were then taken and differential counts made.

TABLE 2 Absolute Leukocyte Mean Percent Count in BALF Eosinophils inBALF Sham 15.4 × 10⁴ ± 1.72 × 10⁴ 20.33% ± 7.1%  Particulate   5 × 10⁴ ±1.18 × 10⁴  3.2% ± 2.9%, 1 mg Particulate 11.14 × 10⁴ ± 1.8 × 10⁴  1.3%± 0.5% 6 mg Soluble 1 mg  3.8 × 10⁴ ± 0.37 × 10⁴ 2.1% ± 1.5% Soluble 6mg   4 × 10⁴ ± 1.5 × 10⁴ 6.2% ± 4.2% Naïve 2.2 × 10⁴ ± 0.2 × 10⁴  0.1% ±0.01%

Absolute leukocyte counts in BALF were increased in sham treated mice(15.4×10⁴±1.72×10⁴ P<0.001) compared to naïve mice (2.2×10⁴±0.2×10⁴P<0.001). Absolute leukocyte counts in BALF were decreased in alltreated groups compared to sham (particulate 1 mg 5×10⁴±1,18×10⁴,P<0.001; particulate 6 mg 11.4×10⁴±1.8×10⁴, P<0.001; soluble 1 mg3.8×104±0.37×10⁴, P<0.001; soluble 6 mg 4×10⁴±1.5×10⁴<P0.001). The meanpercent eosinophils in BALF of sham treated mice (20,33%±7.1%, P<0.008)were elevated compared to naïve (0.1%±0.01%, P<0.008). The mean percenteosinophils in BALF of treated mice were decreased compared to shamtreated mice (particulate 1 mg 3.2%±2.9%, P=0.011; particulate 6 mg1.3%±0,5% P=0.006; soluble 6 mg 6,2%±4.2%, P=0.17).

Example 2

Five week-old AKR mice were pre-treated with oral β-glucan daily for 5consecutive days weekly, for 4 weeks prior to sensitization, particulate1 mg, particulate 6 mg, water-soluble 1 mg, or water-soluble 6 mg. Theycontinued treatment throughout the sensitization and challenge phase.Sham treated mice received an equivalent volume of water. Naïve miceserved as negative controls. β-glucan treated mice and sham treated micewere then intraperitoneally sensitized to ovalbumin (OVA) and aluminiumhydroxide twice at a 7 day interval, followed by 4 sets of intratrachealchallenges at 7-10 day intervals with OVA. Airway pressure time index(APTI) was recorded. 72 hours following the last i.t. challenge, airwayresponsiveness was determined by measuring airway pressure changesfollowing intravenous (i.v.) acetylcholine (Ach) challenge. Briefly,mice were anaesthetized with sodium pentobarbital (80 mg/kg), andventilated via a tracheal cannula (18 gauge) at the rate of 120 perminute with a consistent tidal volume of air (0.2 ml) with RSP 1002Pressure Controlled Respirator System (IKent Scientific Corporation,CT). Muscle paralysis was provided by i.v. administration ofdecamethonium bromide (25 mg/kg). Airway pressure was measured with apressure transducer via a port in the trachea. Two minutes afterestablishing a stable airway pressure recording. Ach was injected i.v.(50 μg/kg). Airway pressure changes were then viewed and recorded, andpulmonary parameters were generated with the software respiratory dataacquisition system VENTP (Kent Scientific Corporation, CT). Thetime-integrated changes in peak airway pressure referred to as theairway pressure-time index (APTI; cm H₂O per second) was calculated andserved as the measurements of airway responsiveness.

TABLE 3 APTI Sham 517.98 ± 27.65 Particulate 1 mg 267.86 ± 27.80Particulate 6 mg 284.36 ± 18.76 Soluble 1 mg 210.46 ± 13.92 Soluble 6 mg524.25 ± 98.35 Naïve 145.74 ± 24.76

APTI was increased in sham-treated mice (517.98±27.55, P<0.001) comparedto naïve mice (145.74±24.76, P<0.001). All β-glucan-treated mice(particulate 1 mg 267.86±27.80, P<0.001; particulate 6 mg 284.36±18.76,P<0.001; soluble 1 mg 210.46±13.92 P<0.001) had decreased APTI comparedwith sham treated mice, with exception to water-soluble β-glucan 6 mgtreated mice 524.25±98.35 P1).

Example 3

Five week-old AKR mice were pre-treated with oral β-glucan daily for 5consecutive days weekly, for 4 weeks prior to sensitization, particulate1 mg, particulate 6 mg, water-soluble 1 mg, or water-soluble 6 mg. Theycontinued treatment throughout the sensitization and challenge phase.Sham treated mice received an equivalent volume of water. Naïve miceserved as negative controls. β-glucan treated mice and sham treated micewere then intraperitoneally sensitized to ovalbumin (OVA) and aluminiumhydroxide twice at a 7 day interval, followed by 4 sets of intratrachealchallenges at 7-10 day intervals with OVA.

Murine splenocytes were cultured with ovalbumin (OVA) and thesupernatants were assessed for cytokine levels by Elisa. Splenocytesfrom mice treated with 1 mg water soluble SBG had decreased productionof IL5 compared with sham treated mice without SBG (199,67 pg/ml versus297,86 pg/ml, p<0.01) Splenocytes from mice treated with 1 mg granularNBG had decreased IL5 production as compared with sham treated micestimulated with OVA (134,85 pg/ml versus 297.86 pg/ml, p<0.01).Similarly, splenocytes from mice treated with both two beta-glucanpreparations showed reduced production of IL-13 as compared tosplenocytes from sham treated mice.

Example 4

Five week-old AKR mice were pre-treated with oral β-glucan daily for 5consecutive days weekly, for 4 weeks prior to sensitization, particulate1 mg, particulate 6 mg, water-soluble 1 mg, or water-soluble 6 mg. Theycontinued treatment throughout the sensitization and challenge phase.Sham treated mice received an equivalent volume of water. Naïve miceserved as negative controls. β-glucan treated mice and sham treated micewere then intraperitoneally sensitized to ovalbumin (OVA) and aluminiumhydroxide twice at a 7 day interval, followed by 4 sets of intratrachealchallenges at 7-10 day intervals with OVA.

Murine splenocytes were cultured with ovalbumin (OVA) and the culturesupernatants were collected after 72 hour culture and assessed forcytokine levels by Elisa. The cytokine levels to OVA stimulation werenormalized by subtracting the respective cyokine levels from mediumalone.

As compared to the sham-treated group, P-BG and S-BG reduced IL-4 by 82%and 36% respectively reduced IL-5 by 56% and 54% respectively andreduced IL-13 by 81% and 96% respectively (FIG. 11). There were nosignificant differences in IL-10, IFN-γ and TGF-β levels between theP-BG or S-BG treated groups and sham-treated groups (FIG. 11).

Example 5

Five week-old AKR mice were pre-treated with oral β-glucan daily for 5consecutive days weekly, for 4 weeks prior to sensitization, particulate1 mg, particulate 6 mg, water-soluble 1 mg, or water-soluble 6 mg. Theycontinued treatment throughout the sensitization and challenge phase.Sham treated mice received an equivalent volume of water. Naïve miceserved as negative controls. β-glucan treated mice and sham treated micewere then intraperitoneally sensitized to ovalbumin (OVA) and aluminiumhydroxide twice at a 7 day interval, followed by 4 sets of intratrachealchallenges at 7-10 day intervals with OVA.

The effect of BG on cytokine profiles of lungs from different groups ofmice were determined by evaluating cytokine mRNA levels usingquantitative real-time PCR (FIG. 12). IL-4, IL-5 and IL-13 mRNA levelswere increased 3, 1 and 7 fold respectively in the sham treated groupcompared to the naïve group indicating a Th2 mediated response. Incontrast, in both of the BG-treated groups, the Th2 mRNA levels, inparticular Il-4 and IL-13 levels, were reduced compared with the shamtreated group. Il-4 was reduced by 55% and 48% in the P-BG and S-BGgroups respectively. IL-13 was reduced by 40% and 53% in the P-BG andS-BG groups respectively. However, there were no differences in IL-10,IFN-γ and TGF-β transcript levels between the treated groups and thesham treated group.

Example 6

Five week -old AKR mice were pre-treated with oral β-glucan daily for 5consecutive days weekly, for 4 weeks prior to sensitization, particulate1 mg, particulate 6 mg, water-soluble 1 mg, or water-soluble 6 mg. Theycontinued treatment throughout the sensitization and challenge phase.Sham treated mice received an equivalent volume of water. Naïve miceserved as negative controls. β-glucan treated mice and sham treated micewere then intraperitoneally sensitized to ovalbumin (OVA) and aluminiumhydroxide twice at a 7 day interval, followed by 4 sets of intratrachealchallenges at 7-10 day intervals with OVA.

Approximately 100 μl of blood was obtained via retro-orbital punctureimmediately prior to APTI. Plasma antigen-specific IgE, IgG1 and IgG2alevels were determined by ELISA. There was no statistically significantdifference in OVA-specific IgE, IgG1 or IgG2a levels between the BGtreated and sham treated groups (FIG. 13).

The above examples demonstrate that both NBG treatment and SBG treatmentsignificantly reduce the allergic pulmonary inflammation typical ofbronchial asthma, and reduces significantly the airway airflowresistance typical of bronchial asthma.

Increased airway resistance (hyperactivity) is the defining element ofbronchial asthma and related diseases. Our results demonstrate that oraladministration of water soluble or granular (particulate) β1-3 glucanreduces hyperactivity and thus appears to be an effectivetreatment/prophylaxis for asthmatic conditions, probably by shifting animmune reaction in the lung from a Th₂ type in the direction of a Th₁,type. The exact mechanism is not established. It is known, however, thatβ1-3 glucan stimulates cells of the mononuclear phagocyte lineage toproduce a series of cytokines that may influence lymphocyte productionof Il-4 and IFN-γ, pivotal for the Th₁/Th₂ balance.

The granular/particulate β1-3 glucan was effective at both dosagestested and suggests an expected dose-effect relationship. The solubleβ1-3 glucan, however, had a stronger effect at the lower dose—which maysuggest a narrower effective dose range, or stimulation of inhibitorysubstances at higher concentrations.

The Examples show that S-BG and P-BG treatment suppressed Th2 responseswith no significant effect on Th1 or T regulatory (IL-10) or cytokine-3(TGF-β). This finding suggests that the prophylactic effect of both S-BGand P-BG on AHR and pulmonary inflammation is associated withdown-regulation of Th2 responses. However, BG had no statisticallysignificant effect on the OVA-specific immunoglobulins, IgE, IgG1 orIgG2a which indicates that the majority of the effect of β-glucan is onT-cells.

Example 7

BBG (the same glucan product as NBG defined herein) was tested in thefollowing scenarios

A: Inhibitory Effect of BBG on Nasal Mucus Secretion

METHOD: Toluene-2,4-diisocynate(TDI)-sensitized guinea pigs were used asan allergic rhinitis model. The amount of nasal mucus was measured afteroral administration of BBG.RESULT & CONCLUSION: The amount of nasal mucus of BBG-ingested group was66.7% lower than that of the control group (FIG. 2).

B: Improvement of Symptoms of Cedar Pollen-Induced Allergic Rhinitis

METHOD: BBG had been administered to cedar pollen-sensitized beaglesduring 57 days. IgE levels in blood and intracutaneous reaction wereexamined.RESULT & CONCLUSIONS: IgE levels in blood were reduced duringadministration (FIG. 3). Approximately 30% reduction of average scoreswas admitted after 56 days (FIG. 4), approximately 60% reduction after168 days. The results indicate improvement of allergic reaction withBBG.

Example 8

A study on the effect of the glucan (NBG) towards hay fever wasconducted. In the study, 43 probants were given 500 mg of theparticulate glucan product. The probants took 125 mg of the product 4times a day (in total 500 mg/day) p.o. Half the study population wasgiven the product for the first 18 days (Group A), whereas the remainingtook the product for 37 days (Group B). Based on self-evaluation (asubjective measurement resulting in “scores”) of a number of 4parameters being an evaluation of the condition of the nasal mucosa, thefrequency of sneezing, the effect on eye itching and nasal breathingability; as well as 5 more general parameters like i.e. physicalcondition, appetite, sleep, there was a marked difference between thetwo groups. Overall, about half of the total study population (i.e. bothgroups combined) claimed effect. The scores obtained in that study werea summary of all parameters, meaning a score of 1.00 indicates that theprobant is still afflicted by the symptoms, while a score of 0.00 showsthat the probants have no further symptoms after taking the glucanproduct.

The results of these studies are presented in FIGS. 5-10.

1. A method of treating asthma or a related disease of abnormal pulmonary function in an animal comprising administering to said animal an effective amount of a glucan having a beta-(1,3)-backbone with one or more beta-(1,3)-side chains linked thereto.
 2. The method according to claim 1 wherein the source of the glucan is selected from the group consisting of yeast, mushrooms or other fungi, Gramineae, algae and bacteria.
 3. The method according to claim 2 wherein the yeast is Saccharomyces cerevisiae.
 4. The method according to claim 1 wherein the glucan is at least partially isolated from other cell wall components.
 5. The method according to claim 1 wherein the glucan has a chemical structure which is altered as compared to its naturally occurring structure.
 6. The method according to claim 1 wherein the glucan is underivatized by chemical groups.
 7. The method according to claim 1 wherein the glucan is essentially free of repetitive beta-(1,6)-linked glucosyl units.
 8. The method according to claim 1 wherein the glucan is in particulate form.
 9. The method according to claim 1 wherein the glucan is solubilized.
 10. The method according to claim 8 wherein the molecular weight of the glucan is in the range of 150 kDa to 300 kDa.
 11. The method according to claim 9 wherein the molecular weight of the glucan is in the range of about 6 kDa to 30 kDA.
 12. The method according to claim 1 wherein the animal is a human.
 13. The method according to claim 1 wherein the glucan is administered orally.
 14. The method according to claim 13 wherein the glucan is administered in a dose range of 10 to 500 mg per kg body weight per day.
 15. The method according to claim 1 wherein the asthma is bronchial asthma.
 16. The method according to claim 1 wherein the related disease of abnormal pulmonary function is selected from the group consisting of allergic bronchopulmonary aspergillosis, chronic bronchitis with an allergic element and emphysema.
 17. (canceled)
 18. A kit or an administration device comprising a glucan as defined in any one of the preceding claims and information material which describes administering the glucan to a human or other animal.
 19. An inhaler, nebuliser of dry powder device containing a glucan as defined in claim
 1. 20. A product comprising (a) a glucan having a beta-(1,3)-backbone with one or more beta-(1,3)-side chains linked thereto and (b) a second active agent for the treatment of asthma or related diseases of abnormal pulmonary function, as a combined preparation for simultaneous, separate or sequential use in the treatment of asthma or related diseases of abnormal pulmonary function. 